Thiocarbamate inhibitors of alpha-4 integrins

ABSTRACT

The invention provides novel thiocarbamate alpha-4 inhibitors of the general formula (I) that are resistant to metabolism having improved half-life and/or clearance properties compared to corresponding carbamate compounds wherein substituents L, X, Y, Z, R 1 –R 4 , m, n, o and p are as defined herein. Also provided are compositions comprising compounds of formula I and a carrier, diluent or excipient as well as methods of treating a disease or condition mediated by the binding interaction of alpha-4 integrins to its ligands such as inflammatory diseases.

This is a continuation application filed under 37 CFR 1.53(b) ofapplication Ser. No. 10/850,002 filed May 19, 2004, now U.S. Pat. No.7,015,323, which claims priority to provisional application Ser. No.60/472,072 filed on May 20, 2003, each of which is incorporated hereinby reference.

FIELD OF THE INVENTION

The present invention relates to organic compounds useful for therapyand/or prophylaxis in a mammal, in particular tyrosine analogs treatingconditions mediated by alpha-4 integrins.

BACKGROUND OF THE INVENTION

The integrins are α/β heterodimeric cell surface receptors involved innumerous cellular processes from cell adhesion to gene regulation.Hynes, R. O., Cell, 1992, 69:11–25; Hemler, M. E., Annu. Rev. Immunol.,1990, 8:365–368. Several integrins have been implicated in diseaseprocesses and have generated widespread interest as potential targetsfor drug discovery. Sharar, S. R. et al., Springer Semin. Immunopathol.,1995, 16:359–378. In the immune system integrins are involved inleukocyte trafficking, adhesion and infiltration during inflammatoryprocesses. Nakajima, H. et al., J. Exp. Med., 1994, 179:1145–1154.Differential expression of integrins regulates the adhesive propertiesof cells and different integrins are involved in different inflammatoryresponses. Butcher, E. C. et al., Science, 1996, 272:60–66. The alpha4integrins (i.e. alpha4beta1 (α4β1) and alpha4beta7 (α4β-7)) areexpressed primarily on monocytes, lymphocytes, eosinophils, basophils,and macrophages but not on neutrophils. Elices, M. J. et al., Cell,1990, 60:577–584. The primary ligands for α4 integrins are theendothelial surface proteins mucosal addressin cell adhesion molecule(MAdCAM) and vascular cell adhesion molecule (VCAM) with lower affinity.Makarem, R. et al., J. Biol. Chem., 1994, 269:4005–4011. The binding ofthe α4β7 or α4β1 to MAdCAM and/or VCAM expressed on high endothelialvenules (HEVs) at sites of inflammation results in firm adhesion of theleukocyte to the endothelium followed by extravasation into the inflamedtissue. Chuluyan, H. E. et al., Springer Semin. Immunopathol., 1995,16:391–404.

Monoclonal antibodies directed against α4β1, α4β7, MAdCAM or VCAM havebeen shown to be effective modulators in animal models of chronicinflammatory diseases such as asthma (Laberge, S. et al., Am. J. Respir.Crit. Care Med., 1995, 151:822–829.), rheumatoid arthritis (RA;Barbadillo, C. et al., Springer Semin. Immunopathol., 1995, 16:375–379),colitis (Viney et al, J. Immunol., 1996, 157: 2488–2497) andinflammatory bowel diseases (IBD; Podalski, D. K., N. Eng. J. Med.,1991, 325:928–937; Powrie, F. et al., Ther. Immunol., 1995, 2:115–123).While antibodies can be effective inhibitors of alpha-4 integrins, theyare inherently difficult and expensive to manufacture. They are also notorally bioavailable and inconveniently require administration byinjection from a physician or other qualified healthcare giver.

In an attempt to find more convenient treatments, many types of smallmolecules have been made to inhibit binding interaction of alpha-4integrins with their ligands, a promising example of which arephenylalanine derivatives such as those described in U.S. Pat. No.6,410,781, U.S. Pat. No. 6,229,011, U.S. Pat. No. 6,329,372, EP1,270,547, WO 01/68,586 and WO 99/36,393. A particular type of potentalpha-4 integrin inhibitor are tyrosine compounds described in U.S. Pat.No. 6,469,047 which are derivatized at the hydroxyl group to form acarbamate. A representative compound disclosed in U.S. Pat. No.6,469,047 is incorporates a

tyrosine residue conjugated to a morpholino heterocycle by way of acarbamate linkage. The carbamates are potent inhibitors of alpha-4integrins but have been shown to metabolize rapidly in vivo yielding aphenoxy metabolite and thereby having a short half life.

Accordingly, there remains a need for small molecule inhibitors ofalpha-4 integrins that are resistant to metabolism having prolonged invivo half-life.

SUMMARY OF THE INVENTION

In one aspect of the present invention there is provided novelthiocarbamate alpha-4 inhibitors of formula (I) that are resistant tometabolism having improved half-life and/or clearance propertiescompared to corresponding carbamate compounds:

wherein

-   q is 0 or 1;-   T is O, CHR⁶, NR⁶, S, SO, SO₂, —NR⁶C(O)—, —C(O)NR⁶—;-   R^(a) and R^(b) are each independently hydrogen, alkyl, alkoxy, a    carbocycle, a heterocycle, optionally substituted with halogen,    hydroxy, amino, carboxyl, nitro, cyano, a carbocycle or a    heterocycle; and one to three carbon atoms of said alkyl and alkoxy    groups are optionally replaced with carbonyl, NR⁶, O, S, SO or SO₂;    or R^(a) and R^(b) together with the nitrogen to which they are    attached may form a heterocycle or heteroaryl group substituted with    0–4 R¹ substituents;-   R^(c) is H, alkyl, optionally substituted with hydroxy, halogen,    alkoxy, amino, a carbocycle or a heterocycle; and a carbon atom of    said alkyl is optionally replaced with carbonyl, NR⁶, O, S, SO or    SO₂;-   L is —C(S)—O— or —C(O)—S—;-   X is O, NR⁵, CR¹R⁶, S, SO or SO₂;-   Y is CH₂ or absent when p is 0;-   Z is H or lower alkyl, or when p is 1 then Z and Y together with the    atoms from which they depend form a 5 member saturated or partially    unsaturated 5 or 6 member heterocycle;-   R¹ in each instance is independently selected from the group    consisting of hydroxy, amino, amidine, guanidine, carboxyl, nitro,    cyano, thiol, alkyl, alkoxy a carbocycle and a heterocycle wherein    said alkyl and alkoxy groups are optionally substituted with one or    more hydroxyl, halogen, amino, amidine, guanidine, carboxyl, nitro,    cyano, carbocycle or heterocycle; and one to three carbon atoms of    said alkyl and alkoxy groups are optionally replaced with carbonyl,    N R⁶, O, S, SO or SO₂; and said carbocycle and heterocycle group is    optionally substituted with one or more hydroxyl, halogen, amino,    amidine, guanidine carboxyl, nitro, cyano, alkyl, alkoxy or    haloalkyl;    -   or two R¹ substituents together with the atoms from which they        depend form a fused or bridged heterocycle optionally        substituted with one or more hydroxyl, halogen, amino, amidine,        guanidine carboxyl, nitro, cyano, alky, alkoxy or haloalkyl;-   R² and R³ in each instance are independently selected from the group    consisting of hydroxy, amino, amidine, guanidine, carboxyl, nitro,    cyano, thiol, alkyl, alkoxy, a carbocycle and a heterocycle wherein    said alkyl and alkoxy groups are optionally substituted with one or    more hydroxyl, halogen, amino, amidine, guanidine, carboxyl, nitro,    cyano, alkoxy, carbocycle or heterocycle; and one to three carbon    atoms of said alkyl group is optionally replaced with carbonyl, NR⁶,    O, S, SO or SO₂; and said carbocycle and heterocycle group is    optionally substituted with one or more hydroxyl, halogen, amino,    amidine, guanidine, carboxyl, nitro, cyano, alkyl, alkoxy or    haloalkyl;-   R⁴ is H, alkyl, a carbocycle or heterocycle wherein said alkyl is    optionally substituted with a carbocycle or heterocycle and said    alkyl, carbocycle and heterocycle are optionally substituted with    lower alkyl, halogen, hydroxyl, alkoxy, haloalkyl or amino;-   R⁵ in each instance is independently selected from the group    consisting of H, alkyl, a carbocycle and a heterocycle wherein said    alkyl group is optionally substituted with one or more hydroxyl,    halogen, amino, amidine, guanidine, carboxyl, nitro, cyano, alkoxy    carbocycle or heterocycle; and one to three carbon atoms of said    alkyl group is optionally replaced with carbonyl, NR⁶, O, S, SO or    SO₂; and said carbocycle and heterocycle group is optionally    substituted with one or more hydroxyl, halogen, amino, amidine,    guanidine carboxyl, nitro, cyano, alkyl, alkoxy or haloalkyl;-   R⁶ in each instance is independently H, alkyl or a carbocycle;-   m, n, and o are each independently 0–4;-   p is 0 or 1; and-   salts and solvates thereof.

In another aspect of the invention, there are provided compositionscomprising compounds of formula I and a carrier, diluent or excipient.

In another aspect of the invention, there are provided methods oftreating a disease or condition mediated by the binding interaction ofalpha-4 integrins to its ligands, comprising administering to a mammalan effective amount of the compound of formula I.

DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

“Alkyl” means a branched or unbranched, saturated or unsaturated (i.e.alkenyl, alkynyl) aliphatic hydrocarbon group, having up to 12 carbonatoms unless otherwise specified. When used as part of another term, forexample “alkylamino”, the alkyl portion is preferably a saturatedhydrocarbon chain, however also includes unsaturated hydrocarbon carbonchains such as “alkenylamino” and “alkynylamino. Examples of alkylgroups include methyl, ethyl, n-propyl, isopropyl, n-butyl, iso-butyl,sec-butyl, tert-butyl, n-pentyl, 2-methylbutyl, 2,2-dimethylpropyl,n-hexyl, 2-methylpentyl, 2,2-dimethylbutyl, n-heptyl, 3-heptyl,2-methylhexyl, and the like. The terms “lower alkyl” “C₁–C₄ alkyl” and“alkyl of 1 to 4 carbon atoms” are synonymous and used interchangeablyto mean methyl, ethyl, 1-propyl, isopropyl, cyclopropyl, 1-butyl,sec-butyl or t-butyl. Unless specified, substituted, alkyl groups maycontain one (preferably), two, three or four substituents which may bethe same or different. Examples of the above substituted alkyl groupsinclude, but are not limited to; cyanomethyl, nitromethyl,hydroxymethyl, trityloxymethyl, propionyloxymethyl, aminomethyl,carboxymethyl, carboxyethyl, carboxypropyl, alkyloxycarbonylmethyl,allyloxycarbonylaminomethyl, carbamoyloxymethyl, methoxymethyl,ethoxymethyl, t-butoxymethyl, acetoxymethyl, chloromethyl, bromomethyl,iodomethyl, trifluoromethyl, 6-hydroxyhexyl, 2,4-dichloro(n-butyl),2-amino(iso-propyl), 2-carbamoyloxyethyl and the like. The alkyl groupmay also be substituted with a carbocycle group. Examples includecyclopropylmethyl, cyclobutylmethyl, cyclopentylmethyl, andcyclohexylmethyl groups, as well as the corresponding-ethyl, -propyl,-butyl, -pentyl, -hexyl groups, etc. Preferred substituted alkyls aresubstituted methyls e.g. a methyl group substituted by the samesubstituents as the “substituted C_(n)–C_(m) alkyl” group. Examples ofthe substituted methyl group include groups such as hydroxymethyl,protected hydroxymethyl (e.g. tetrahydropyranyloxymethyl),acetoxymethyl, carbamoyloxymethyl, trifluoromethyl, chloromethyl,carboxymethyl, bromomethyl and iodomethyl.

“Amidine” denotes the group —C(NH)—NHR wherein R is H or alkyl oraralkyl. A preferred amidine is the group —NH—C(NH)—NH₂.

“Amino” denotes primary (i.e. —NH₂), secondary (i.e. —NRH) and tertiary(i.e. —NRR) amines. Preferred secondary and tertiary amines arealkylamine, dialkylamine, arylamine, diarylamine, aralkylamine anddiaralkylamine. Particular preferred secondary and tertiary amines areas methylamine, ethylamine, propylamine, isopropylamine, phenylamine,benzylamine dimethylamine, diethylamine, dipropylamine anddisopropylamine. The term “amidine” denotes the group —C(NH)NHR whereinR is H or alkyl or aralkyl. Preferred amidine is the group —C(NH)NH₂.

“Amino-protecting group” as used herein refers to a derivative of thegroups commonly employed to block or protect an amino group whilereactions are carried out on other functional groups on the compound.Examples of such protecting groups include carbamates, amides, alkyl andaryl groups, imines, as well as many N-heteroatom derivatives which canbe removed to regenerate the desired amine group. Further examples ofthese groups are found in T. W. Greene and P. G. M. Wuts, “ProtectiveGroups in Organic Synthesis”, 2nd ed., John Wiley & Sons, Inc., NewYork, N.Y., 1991, chapter 7; E. Haslam, “Protective Groups in OrganicChemistry”, J. G. W. McOmie, Ed., Plenum Press, New York, N.Y., 1973,Chapter 5, and T. W. Greene, “Protective Groups in Organic Synthesis”,John Wiley and Sons, New York, N.Y., 1981. The term “protected amino”refers to an amino group substituted with one of the aboveamino-protecting groups.

“Aryl” when used alone or as part of another term means a carbocyclicaromatic group whether or not fused having the number of carbon atomsdesignated or if no number is designated, up to 14 carbon atoms.Preferred aryl groups include phenyl, naphthyl, biphenyl, phenanthrenyl,naphthacenyl, and the like (see e.g. Lang's Handbook of Chemistry (Dean,J. A., ed) 13^(th) ed. Table 7-2 [1985]) and most preferred phenyl.Substituted phenyl or substituted aryl denotes a phenyl group or arylgroup substituted with one, two, three, four or five, preferably 1–2,1–3 or 1–4 substituents chosen, unless otherwise specified, from halogen(F, Cl, Br, I), hydroxy, protected hydroxy, cyano, nitro, alkyl(preferably C₁–C₆ alkyl), alkoxy (preferably C₁–C₆ alkoxy), benzyloxy,carboxy, protected carboxy, carboxymethyl, protected carboxymethyl,hydroxymethyl, protected hydroxymethyl, aminomethyl, protectedaminomethyl, trifluoromethyl, alkylsulfonylamino, arylsulfonylamino,heterocyclylsulfonylamino, heterocyclyl, aryl, or other groupsspecified. One or more methyne (CH) and/or methylene (CH₂) groups inthese substituents may in turn be substituted with a similar group asthose denoted above. Examples of the term “substituted phenyl” includesbut is not limited to a mono- or di(halo)phenyl group such as2-chlorophenyl, 2-bromophenyl, 4-chlorophenyl, 2,6-dichlorophenyl,2,5-dichlorophenyl, 3,4-dichlorophenyl, 3-chlorophenyl, 3-bromophenyl,4-bromophenyl, 3,4-dibromophenyl, 3-chloro-4-fluorophenyl,2-fluorophenyl and the like; a mono- or di(hydroxy)phenyl group such as4-hydroxyphenyl, 3-hydroxyphenyl, 2,4-dihydroxyphenyl, theprotected-hydroxy derivatives thereof and the like; a nitrophenyl groupsuch as 3- or 4-nitrophenyl; a cyanophenyl group, for example,4-cyanophenyl; a mono- or di(lower alkyl)phenyl group such as4-methylphenyl, 2,4-dimethylphenyl, 2-methylphenyl,4-(iso-propyl)phenyl, 4-ethylphenyl, 3-(n-propyl)phenyl and the like; amono or di(alkoxy)phenyl group, for example, 3,4-dimethoxyphenyl,3-methoxy-4-benzyloxyphenyl,3-methoxy-4-(1-chloromethyl)benzyloxy-phenyl, 3-ethoxyphenyl,4-(isopropoxy)phenyl, 4-(t-butoxy)phenyl, 3-ethoxy-4-methoxyphenyl andthe like; 3- or 4-trifluoromethylphenyl; a mono- or dicarboxyphenyl or(protected carboxy)phenyl group such 4-carboxyphenyl, a mono- ordi(hydroxymethyl)phenyl or (protected hydroxymethyl)phenyl such as3-(protected hydroxymethyl)phenyl or 3,4-di(hydroxymethyl)phenyl; amono- or di(aminomethyl)phenyl or (protected aminomethyl)phenyl such as2-(aminomethyl)phenyl or 2,4-(protected aminomethyl)phenyl; or a mono-or di(N-(methylsulfonylamino))phenyl such as3-(N-methylsulfonylamino))phenyl. Also, the term “substituted phenyl”represents disubstituted phenyl groups where the substituents aredifferent, for example, 3-methyl-4-hydroxyphenyl,3-chloro-4-hydroxyphenyl, 2-methoxy-4-bromophenyl,4-ethyl-2-hydroxyphenyl, 3-hydroxy-4-nitrophenyl,2-hydroxy-4-chlorophenyl, and the like, as well as trisubstituted phenylgroups where the substituents are different, for example3-methoxy-4-benzyloxy-6-methyl sulfonylamino,3-methoxy-4-benzyloxy-6-phenyl sulfonylamino, and tetrasubstitutedphenyl groups where the substituents are different such as3-methoxy-4-benzyloxy-5-methyl-6-phenyl sulfonylamino. Preferredsubstituted phenyl groups include the 2-chlorophenyl, 2-aminophenyl,2-bromophenyl, 3-methoxyphenyl, 3-ethoxy-phenyl, 4-benzyloxyphenyl,4-methoxyphenyl, 3-ethoxy-4-benzyloxyphenyl, 3,4-diethoxyphenyl,3-methoxy-4-benzyloxyphenyl,3-methoxy-4-(1-chloromethyl)benzyloxy-phenyl,3-methoxy-4-(1-chloromethyl)benzyloxy-6-methyl sulfonyl aminophenylgroups. Fused aryl rings may also be substituted with any, preferably 1,2 or 3, of the substituents specified herein in the same manner assubstituted alkyl groups.

“Carbocyclyl”, “carbocyclylic”, “carbocycle” and “carbocyclo” alone andwhen used as a moiety in a complex group such as a carbocycloalkylgroup, refers to a mono-, bi-, or tricyclic aliphatic ring having 3 to14 carbon atoms and preferably 3 to 7 carbon atoms which may besaturated or unsaturated, aromatic or non-aromatic. Preferred saturatedcarbocyclic groups include cyclopropyl, cyclobutyl, cyclopentyl andcyclohexyl groups and more preferred are cyclopropyl and cyclohexyl andmost preferred is cyclohexyl. Preferred unsaturated carbocycles arearomatic e.g. aryl groups as previously defined, the most preferredbeing phenyl. The terms “substituted carbocyclyl”, “carbocycle” and“carbocyclo” mean these groups substituted by the same substituents asthe “substituted alkyl” group.

“Carboxy-protecting group” as used herein refers to one of the esterderivatives of the carboxylic acid group commonly employed to block orprotect the carboxylic acid group while reactions are carried out onother functional groups on the compound. Examples of such carboxylicacid protecting groups include 4-nitrobenzyl, 4-methoxybenzyl,3,4-dimethoxybenzyl, 2,4-dimethoxybenzyl, 2,4,6-trimethoxybenzyl,2,4,6-trimethylbenzyl, pentamethylbenzyl, 3,4-methylenedioxybenzyl,benzhydryl, 4,4′-dimethoxybenzhydryl, 2,2′,4,4′-tetramethoxybenzhydryl,alkyl such as t-butyl or t-amyl, trityl, 4-methoxytrityl,4,4′-dimethoxytrityl, 4,4′,4″-trimethoxytrityl, 2-phenylprop-2-yl,trimethylsilyl, t-butyldimethylsilyl, phenacyl, 2,2,2-trichloroethyl,beta-(trimethylsilyl)ethyl, beta-(di(n-butyl)methylsilyl)ethyl,p-toluenesulfonylethyl, 4-nitrobenzylsulfonylethyl, allyl, cinnamyl,1-(trimethylsilylmethyl)prop-1-en-3-yl, and like moieties. The speciesof carboxy-protecting group employed is not critical so long as thederivatized carboxylic acid is stable to the condition of subsequentreaction(s) on other positions of the molecule and can be removed at theappropriate point without disrupting the remainder of the molecule. Inparticular, it is important not to subject a carboxy-protected moleculeto strong nucleophilic bases or reductive conditions employing highlyactivated metal catalysts such as Raney nickel. (Such harsh removalconditions are also to be avoided when removing amino-protecting groupsand hydroxy-protecting groups, discussed below.) Preferred carboxylicacid protecting groups are the allyl and p-nitrobenzyl groups. Similarcarboxy-protecting groups used in the cephalosporin, penicillin andpeptide arts can also be used to protect a carboxy group substituents.Further examples of these groups are found in T. W. Greene and P. G. M.Wuts, “Protective Groups in Organic Synthesis”, 2nd ed., John Wiley &Sons, Inc., New York, N.Y., 1991, chapter 5; E. Haslam, “ProtectiveGroups in Organic Chemistry”, J. G. W. McOmie, Ed., Plenum Press, NewYork, N.Y., 1973, Chapter 5, and T. W. Greene, “Protective Groups inOrganic Synthesis”, John Wiley and Sons, New York, N.Y., 1981, Chapter5. The term “protected carboxy” refers to a carboxy group substitutedwith one of the above carboxy-protecting groups.

“Guanidine” denotes the group —NH—C(NH)—NHR wherein R is H or alkyl oraralkyl. Preferred guanidine is the group —NH—C(NH)—NH₂.

“Hydroxy-protecting group” as used herein refers to a derivative of thehydroxy group commonly employed to block or protect the hydroxy groupwhite reactions are carried out on other functional groups on thecompound. Examples of such protecting groups includetetrahydropyranyloxy, acetoxy, carbamoyloxy, trifluoro, chloro, carboxy,bromo and iodo groups. Further examples of these groups are found in T.W. Greene and P. G. M. Wuts, “Protective Groups in Organic Synthesis”,2nd ed., John Wiley & Sons, Inc., New York, N.Y., 1991, chapters 2–3; E.Haslam, “Protective Groups in Organic Chemistry”, J. G. W. McOmie, Ed.,Plenum Press, New York, N.Y., 1973, Chapter 5, and T. W. Greene,“Protective Groups in Organic Synthesis”, John Wiley and Sons, New York,N.Y., 1981. The term “protected hydroxy” refers to a hydroxy groupsubstituted with one of the above hydroxy-protecting groups.

“Heterocyclic group”, “heterocyclic”, “heterocycle”, “heterocyclyl”, or“heterocyclo” alone and when used as a moiety in a complex group such asa heterocycloalkyl group, are used interchangeably and refer to anymono-, bi-, or tricyclic, saturated or unsaturated, aromatic(heteroaryl) or non-aromatic ring having the number of atoms designated,generally from 5 to about 14 ring atoms, where the ring atoms are carbonand at least one heteroatom (nitrogen, sulfur or oxygen) and preferably1 to 4 heteroatoms. Typically, a 5-membered ring has 0 to 2 double bondsand 6- or 7-membered ring has 0 to 3 double bonds and the nitrogen orsulfur heteroatoms may optionally be oxidized (e.g. SO, SO₂), and anynitrogen heteroatom may optionally be quaternized. Preferrednon-aromatic heterocycles include morpholinyl (morpholino),pyrrolidinyl, oxiranyl, oxetanyl, tetrahydrofuranyl, 2,3-dihydrofuranyl,2H-pyranyl, tetrahydropyranyl, thiiranyl, thietanyl,tetrahydrothietanyl, aziridinyl, azetidinyl, 1-methyl-2-pyrrolyl,piperazinyl and piperidinyl. A “heterocycloalkyl” group is a heterocyclegroup as defined above covalently bonded to an alkyl group as definedabove. Preferred 5-membered heterocycles containing a sulfur or oxygenatom and one to three nitrogen atoms include thiazolyl, in particularthiazol-2-yl and thiazol-2-yl N-oxide, thiadiazolyl, in particular1,3,4-thiadiazol-5-yl and 1,2,4-thiadiazol-5-yl, oxazolyl, preferablyoxazol-2-yl, and oxadiazolyl, such as 1,3,4-oxadiazol-5-yl, and1,2,4-oxadiazol-5-yl. Preferred 5-membered ring heterocycles containing2 to 4 nitrogen atoms include imidazolyl, preferably imidazol-2-yl;triazolyl, preferably 1,3,4-triazol-5-yl; 1,2,3-triazol-5-yl,1,2,4-triazol-5-yl, and tetrazolyl, preferably 1H-tetrazol-5-yl.Preferred benzo-fused 5-membered heterocycles are benzoxazol-2-yl,benzthiazol-2-yl and benzimidazol-2-yl. Preferred 6-memberedheterocycles contain one to three nitrogen atoms and optionally a sulfuror oxygen atom, for exampe pyridyl, such as pyrid-2-yl, pyrid-3-yl, andpyrid-4-yl; pyrimidyl, preferably pyrimid-2-yl and pyrimid-4-yl;triazinyl, preferably 1,3,4-triazin-2-yl and 1,3,5-triazin-4-yl;pyridazinyl, in particular pyridazin-3-yl, and pyrazinyl. The pyridineN-oxides and pyridazine N-oxides and the pyridyl, pyrimid-2-yl,pyrimid-4-yl, pyridazinyl and the 1,3,4-triazin-2-yl groups, are apreferred group. Substituents for optionally substituted heterocycles,and further examples of the 5- and 6-membered ring systems discussedabove can be found in W. Druckheimer et al., U.S. Pat. No. 4,278,793.

“Heteroaryl” alone and when used as a moiety in a complex group such asa heteroaralkyl group, refers to any mono-, bi-, or tricyclic aromaticring system having the number of atoms designated where at least onering is a 5-, 6- or 7-membered ring containing from one to fourheteroatoms selected from the group nitrogen, oxygen, and sulfur, andpreferably at least one heteroatom is nitrogen (Lang's Handbook ofChemistry, supra). Included in the definition are any bicyclic groupswhere any of the above heteroaryl rings are fused to a benzene ring.Heteroaryls in which nitrogen or oxygen is the heteroatom are preferred.The following ring systems are examples of the heteroaryl (whethersubstituted or unsubstituted) groups denoted by the term “heteroaryl”:thienyl, furyl, imidazolyl, pyrazolyl, thiazolyl, isothiazolyl,oxazolyl, isoxazolyl, triazolyl, thiadiazolyl, oxadiazolyl, tetrazolyl,thiatriazolyl, oxatriazolyl, pyridyl, pyrimidyl, pyrazinyl, pyridazinyl,thiazinyl, oxazinyl, triazinyl, thiadiazinyl, oxadiazinyl, dithiazinyl,dioxazinyl, oxathiazinyl, tetrazinyl, thiatriazinyl, oxatriazinyl,dithiadiazinyl, imidazolinyl, dihydropyrimidyl, tetrahydropyrimidyl,tetrazolo[1,5-b]pyridazinyl and purinyl, as well as benzo-fusedderivatives, for example benzoxazolyl, benzofuryl, benzothiazolyl,benzothiadiazolyl, benzotriazolyl, benzoimidazolyl and indolyl. Aparticularly preferred group of “heteroaryl” include; 1,3-thiazol-2-yl,4-(carboxymethyl)-5-methyl-1,3-thiazol-2-yl,4-(carboxymethyl)-5-methyl-1,3-thiazol-2-yl sodium salt,1,2,4-thiadiazol-5-yl, 3-methyl-1,2,4-thiadiazol-5-yl,1,3,4-triazol-5-yl, 2-methyl-1,3,4-triazol-5-yl,2-hydroxy-1,3,4-triazol-5-yl, 2-carboxy-4-methyl-1,3,4-triazol-5-ylsodium salt, 2-carboxy-4-methyl-1,3,4-triazol-5-yl, 1,3-oxazol-2-yl,1,3,4-oxadiazol-5-yl, 2-methyl-1,3,4-oxadiazol-5-yl,2-(hydroxymethyl)-1,3,4-oxadiazol-5-yl, 1,2,4-oxadiazol-5-yl,1,3,4-thiadiazol-5-yl, 2-thiol-1,3,4-thiadiazol-5-yl,2-(methylthio)-1,3,4-thiadiazol-5-yl, 2-amino-1,3,4-thiadiazol-5-yl,1H-tetrazol-5-yl, 1-methyl-1H-tetrazol-5-yl,1-(1-(dimethylamino)eth-2-yl)-1H-tetrazol-5-yl,1-(carboxymethyl)-1H-tetrazol-5-yl, 1-(carboxymethyl)-1H-tetrazol-5-ylsodium salt, 1-(methylsulfonic acid)-1H-tetrazol-5-yl, 1-(methylsulfonicacid)-1H-tetrazol-5-yl sodium salt, 2-methyl-1H-tetrazol-5-yl,1,2,3-triazol-5-yl, 1-methyl-1,2,3-triazol-5-yl,2-methyl-1,2,3-triazol-5-yl, 4-methyl-1,2,3-triazol-5-yl, pyrid-2-ylN-oxide, 6-methoxy-2-(n-oxide)-pyridaz-3-yl, 6-hydroxypyridaz-3-yl,1-methylpyrid-2-yl, 1-methylpyrid-4-yl, 2-hydroxypyrimid-4-yl,1,4,5,6-tetrahydro-5,6-dioxo-4-methyl-as-triazin-3-yl,1,4,5,6-tetrahydro-4-(formylmethyl)-5,6-dioxo-as-triazin-3-yl,2,5-dihydro-5-oxo-6-hydroxy-astriazin-3-yl,2,5-dihydro-5-oxo-6-hydroxy-as-triazin-3-yl sodium salt,2,5-dihydro-5-oxo-6-hydroxy-2-methyl-astriazin-3-yl sodium salt,2,5-dihydro-5-oxo-6-hydroxy-2-methyl-as-triazin-3-yl,2,5-dihydro-5-oxo-6-methoxy-2-methyl-as-triazin-3-yl,2,5-dihydro-5-oxo-as-triazin-3-yl,2,5-dihydro-5-oxo-2-methyl-as-triazin-3-yl,2,5-dihydro-5-oxo-2,6-dimethyl-as-triazin-3-yl,tetrazolo[1,5-b]pyridazin-6-yl and8-aminotetrazolo[1,5-b]-pyridazin-6-yl. An alternative group of“heteroaryl” includes; 4-(carboxymethyl)-5-methyl-1,3-thiazol-2-yl,4-(carboxymethyl)-5-methyl-1,3-thiazol-2-yl sodium salt,1,3,4-triazol-5-yl, 2-methyl-1,3,4-triazol-5-yl, 1H-tetrazol-5-yl,1-methyl-1H-tetrazol-5-yl,1-(1-(dimethylamino)eth-2-yl)-1H-tetrazol-5-yl,1-(carboxymethyl)-1H-tetrazol-5-yl, 1-(carboxymethyl)-1H-tetrazol-5-ylsodium salt, 1-(methylsulfonic acid)-H-tetrazol-5-yl, I-(methylsulfonicacid)-1H-tetrazol-5-yl sodium salt, 1,2,3-triazol-5-yl,1,4,5,6-tetrahydro-5,6-dioxo-4-methyl-as-triazin-3-yl,1,4,5,6-tetrahydro-4-(2-formylmethyl)-5,6-dioxo-as-triazin-3-yl,2,5-dihydro-5-oxo-6-hydroxy-2-methyl-as-triazin-3-yl sodium salt,2,5-dihydro-5-oxo-6-hydroxy-2-methyl-as-triazin-3-yl,tetrazolo[1,5-b]pyridazin-6-yl, and8-aminotetrazolo[1,5-b]pyridazin-6-yl.

“Inhibitor” means a compound which reduces or prevents the binding ofalpha4 integrins to their ligands, for example alpha-4beta1 integrin toVCAM-1 ligand alpha-4beta7 integrin to MAdCAM-1 ligand or which reducesor prevents the initiation of a cellular response mediated by thebinding interaction of the alpha-4 integrins to their ligands.

“Pharmaceutically acceptable salts” include both acid and base additionsalts. “Pharmaceutically acceptable acid addition salt” refers to thosesalts which retain the biological effectiveness and properties of thefree bases and which are not biologically or otherwise undesirable,formed with inorganic acids such as hydrochloric acid, hydrobromic acid,sulfuric acid, nitric acid, carbonic acid, phosphoric acid and the like,and organic acids may be selected from aliphatic, cycloaliphatic,aromatic, araliphatic, heterocyclic, carboxylic, and sulfonic classes oforganic acids such as formic acid, acetic acid, propionic acid, glycolicacid, gluconic acid, lactic acid, pyruvic acid, oxalic acid, malic acid,maleic acid, maloneic acid, succinic acid, fumaric acid, tartaric acid,citric acid, aspartic acid, ascorbic acid, glutamic acid, anthranilicacid, benzoic acid, cinnamic acid, mandelic acid, embonic acid,phenylacetic acid, methanesulfonic acid, ethanesulfonic acid,p-toluenesulfonic acid, salicyclic acid and the like.

“Pharmaceutically acceptable base addition salts” include those derivedfrom inorganic bases such as sodium, potassium, lithium, ammonium,calcium, magnesium, iron, zinc, copper, manganese, aluminum salts andthe like. Particularly preferred are the ammonium, potassium, sodium,calcium and magnesium salts. Salts derived from pharmaceuticallyacceptable organic nontoxic bases includes salts of primary, secondary,and tertiary amines, substituted amines including naturally occurringsubstituted amines, cyclic amines and basic ion exchange resins, such asisopropylamine, trimethylamine, diethylamine, triethylamine,tripropylamine, ethanolamine, 2-diethylaminoethanol, trimethamine,dicyclohexylamine, lysine, arginine, histidine, caffeine, procaine,hydrabamine, choline, betaine, ethylenediamine, glucosamine,methylglucamine, theobromine, purines, piperizine, piperidine,N-ethylpiperidine, polyamine resins and the like. Particularly preferredorganic non-toxic bases are isopropylamine, diethylamine, ethanolamine,trimethamine, dicyclohexylamine, choline, and caffeine.

The present invention provides compounds having the general formula I:

wherein q, T R^(a)-R^(c), L, X, Y, Z, R²-R⁴, m, n, o, p are as definedherein.

q is 0 or 1. In a particular embodiment q is 0. In another particularembodiment q is 1.

T is O, CHR⁶, NR⁶, S, SO, SO₂, —NR⁶C(O)—, —C(O)NR⁶—. I a preferredembodiment T is 0 or C(O)NR⁶— and most preferably O.

R^(a) and R^(b) are each independently hydrogen, alkyl, alkoxy, acarbocycle, a heterocycle, optionally substituted with halogen, hydroxy,amino, carboxyl, nitro, cyano, a carbocycle or a heterocycle. One tothree carbon atoms of said alkyl and alkoxy groups are optionallyreplaced with carbonyl, NR⁶, O, S, SO or SO₂ in the same manner asdescribed for R¹. Preferred groups formed by R^(a) and R^(b) with thenitrogen atom from which they depend are:

Alternatively, R^(a) and R^(b) together with the nitrogen to which theyare attached may form a heterocycle group substituted with 0–4 R¹substituents. Preferred heterocycles formed by R^(a) and R^(b) are:

R^(c) is H, alkyl, optionally substituted with hydroxy, halogen, alkoxy,amino, a carbocycle or a heterocycle. One to three carbon atoms of saidalkyl are optionally replaced with carbonyl, NR⁶, O, S, SO or SO₂ in thesame manner as described for R¹ below. In a preferred embodiment R^(c)is an amino acid sidechain, preferably a sidechain of a naturallyoccurring amino acid. In a particularly preferred embodiment R⁶ is H orthe amino acid side chain of glycine, valine, leucine or isoleucine andmost preferably leucine. In a particularly preferred embodiment, q is 1and R^(c) is a leucine side chain while T is O, R³ is t-butyl and ois 1. More preferably q is 1 and R^(c) is a leucine side chain while Tis O, R³ is t-butyl, o is 1, R^(a) and R^(b) form a morpholino ring, Lis —C(S)—O—, Z is H, Y is absent, p is 0 and R⁴ and R⁶ are both H.

L is —C(S)—O— or —C(O)—S— thereby forming a thiocarbamate linkage. In apreferred embodiment L is —C(S)—O—.

Y is CH₂ or is absent when p is 0. In a preferred embodiment p is 0 andY is absent.

Z is H or lower alkyl, or when p is 1 then Z and Y together with theatoms from which they depend form a 5 member saturated or partiallyunsaturated 5 or 6 member heterocycle. In a particular embodiment Z andY are both CH₂ and p is I thereby forming a pyrrolidine ring andconstraining rotation of the tyrosine residue. In a preferred embodimentZ is H or methyl and most preferably H while p is 0 and Y is absent.

R² and R³ in each instance are independently selected from the groupconsisting of hydroxy, amino, amidine, guanidine, carboxyl, nitro,cyano, thiol, alkyl, alkoxy, a carbocycle and a heterocycle. The alkyland alkoxy groups of R² and R³ are optionally substituted with one ormore hydroxyl, halogen, amino, amidine, guanidine, carboxyl, nitro,cyano, alkoxy, carbocycle or heterocycle. Further, one to three carbonatoms of said alkyl group is optionally replaced with carbonyl, NR⁶, O,S, SO or SO₂ in the same manner as described for R¹ below. Thecarbocycle and heterocycle groups of R² and R³ are optionallysubstituted with one or more hydroxyl, halogen, amino, amidine,guanidine, carboxyl, nitro, cyano, alkyl, alkoxy or haloalkyl. In aparticular embodiment, R² is in each instance halogen, alkyl, alkoxy,aryl or aryloxy. In a particularly preferred embodiment, n is 1 and R²is adjacent to the thiocarbamate linkage and is selected from the groupconsisting of Cl, I, methyl, methoxy and phenyl. In another particularlypreferred embodiment n is 0 and R² is absent. In a particularembodiment, R³ is halogen, nitro, carboxyl or alkylsulfonyl optionallysubstituted with halogen, hydroxyl or alkoxy. In another particularembodiment R³ is Cl, nitro, carboxyl, —NHSO₂CF₃, —NHC(O)CF₃ or4-methyl-phenyl. In a particularly preferred embodiment o is 1 and R³ isCl adjacent to the amide linkage.

R⁴ is H, alkyl, a carbocycle or heterocycle wherein said alkyl isoptionally substituted with a carbocycle or heterocycle and said alkyl,carbocycle and heterocycle are optionally substituted with lower alkyl,halogen, hydroxyl, alkoxy, haloalkyl or amino. In a preferred embodimentR⁴ is H, alkyl or aralkyl. In a particularly preferred embodiment R⁴ ismethyl, ethyl, isobutyl or benzyl and more preferably ethyl. In anotherparticularly preferred embodiment R⁴ is H.

R⁶ in each instance is independently H, alkyl or a carbocycle. In apreferred embodiment, R⁶ is H.

and o are each independently 0–4. In a particular embodiment m and n areboth 0–1 and o is 1–2. More preferably, m and n or both 0 and o is 1.

p is 0 or 1. In a preferred embodiment p is 1 and Y is CH₂ therebyforming a heterocycle, more preferably a pyrrolidine. In anotherpreferred p is 0 and Y is absent.

In a particular embodiment, compounds of the invention have the generalformula II

wherein L, Y, Z, R²–R⁴, n, o, p are as defined above and m, X, R¹ and R⁵are as defined herein below.

m is 0–4, preferably 0–2 more preferably 0–1 and most preferably 0. In aparticular embodiment m and o are both 0–1. In another particularembodiment m and n are both 0 and o is 1.

X is O, NR⁵, CR¹R⁶, S, SO or SO₂. In a particular embodiment X is CR¹R⁶.In another particular embodiment X is S. In another embodiment X is SO₂.In a preferred embodiment X is SO. In another preferred embodiment X isNR⁵ wherein R⁵ is as defined below. In a most preferred embodiment X is0 thereby forming a morpholino heterocycle.

R¹ in each instance is independently selected from the group consistingof hydroxy, amino, amidine, guanidine, carboxyl, nitro, cyano, thiol,alkyl, alkoxy a carbocycle and a heterocycle. The carbocycle andheterocycle group of R¹ is optionally substituted with one or more,preferably 1–3, hydroxyl, halogen, amino, amidine, guanidine carboxyl,nitro, cyano, alkyl, alkoxy or haloalkyl. The alkyl and alkoxy groups ofR¹ are optionally substituted with one or more, preferably 1–3,hydroxyl, halogen, amino, amidine, guanidine, carboxyl, nitro, cyano,carbocycle or heterocycle substituents. Further, one to three carbonatoms of said alkyl group (as well as any pending hydrogen atoms, e.g.methylene) are optionally replaced with carbonyl C(O), NR⁶, O, S, SO orSO₂. In a preferred embodiment a carbon atom in an alkyl chain isreplaced to form an alkanoyl, ketone or aldehyde group. In anotherpreferred embodiment, a carbon atom in an alkyl chain is replaced withNR⁶ to form an amine, aminoalkyl or mono- or di-alkylaminoalkyl. Inanother preferred embodiment, a carbon atom is replaced with O to forman alkoxy, alkoxyalkyl (ether) or hydroxyalkyl. In another preferredembodiment, a carbon atom in an alkyl chain is replaced with S to forman alkylthio, thioether or thiolalkyl. In another embodiment, two ormore adjacent carbon atoms in an alkyl chain are replaced with—NR⁶—C(O)—, —C(O)—NR⁶—, —NR⁶—SO, —SO—NR⁶—, —NR⁶—SO₂— or —SO₂—NR⁶—. In apreferred embodiment, two or more carbon atoms in an alkyl chain arereplaced to form amide groups —NR⁶—C(O)-alkyl, —C(O)NR⁶-alkyl; oralkylsulfonyl groups —NR⁶—SO₂-alkyl, —SO₂—NR⁶-alkyl, —N—(SO₂-alkyl)₂ or—SO₂—N(alkyl)₂. Particularly preferred alkylsulfonyl groups are—NH—SO₂-Me, —NH—SO₂-Et, —NH—SO₂—Pr, —NH—SO₂-iPr, —N—(SO₂-Me)₂ and—N—(SO₂-Bu)₂.

In a preferred embodiment R¹ in each instance is H; or alkyl, aryl,heteroaryl each optionally substituted with hydroxyl, halogen amino orcyano. In a particularly preferred embodiment R¹ is H, methyl, ethyl,isopropyl, cyanomethyl. In another particularly preferred embodiment mis 0 and R⁵ is absent. In another particularly preferred embodiment m isI and R¹ is methyl adjacent to the thiocarbamate nitrogen atom.

In another embodiment, two R¹ substituents together with the atoms fromwhich they depend form a fused or bridged heterocycle optionallysubstituted with one or more hydroxyl, halogen, amino, amidine,guanidine carboxyl, nitro, cyano, alky, alkoxy or haloalkyl. A preferredfused heterocycle formed by adjacent R¹ substituents is1,2,3,4-tetrahydroisoquinoline which is optionally substituted with oneor more alkoxy and preferably 6, 7,-dimethoxy substituted. Preferredbridged heterocycles formed by non-adjacent R¹ substituents are2-Oxa-5-azabicyclo[2.2.1]heptane and 2,5-Diazabicyclo[2.2.1]heptane, thelatter optionally N-substituted with an acyl group such as acetyl.

R⁵ in each instance is independently selected from the group consistingof H, alkyl, a carbocycle and a heterocycle. The carbocycle andheterocycle groups of R⁵ are optionally substituted with one or morehydroxyl, halogen, amino, amidine, guanidine carboxyl, nitro, cyano,alkyl, alkoxy or haloalkyl. The alkyl group of R⁵ is optionallysubstituted with one or more hydroxyl, halogen, amino, amidine,guanidine, carboxyl, nitro, cyano, alkoxy, carbocycle or heterocycle.Further, one to three carbon atoms of said alkyl group is optionallyreplaced with carbonyl, NR⁶, O, S, SO or SO₂ in the same manner asdescribed for R¹. In a particular embodiment R⁵ is H, alkyl andalkanoyl, optionally substituted with hydroxyl, halogen, amino or aryl.In a particularly preferred embodiment R⁵ is H, methyl, ethyl or acetyland in a more preferred embodiment H or acetyl.

Compounds of the invention contain one or more asymmetric carbon atoms.Accordingly, the compounds may exist as diastereomers, enantiomers ormixtures thereof. The syntheses of the compounds may employ racemates,diastereomers or enantiomers as starting materials or as intermediates.Diastereomeric compounds may be separated by chromatographic orcrystallization methods. Similarly, enantiomeric mixtures may beseparated using the same techniques or others known in the art. Each ofthe asymmetric carbon atoms may be in the R or S configuration and bothof these configurations are within the scope of the invention.Preferably, compounds of the invention have an S configuration at thealpha carbon of the tyrosine residue or the naturally occurringconfiguration thereof.

The invention also encompasses prodrugs of the compounds describedabove. Suitable prodrugs include known amino-protecting andcarboxy-protecting groups which are released, for example hydrolyzed, toyield the parent compound under physiologic conditions. A preferredclass of prodrugs are compounds in which a nitrogen atom in an amino,amidino, aminoalkyleneamino, iminoalkyleneamino or guanidino group issubstituted with a hydroxy (OH) group, an alkylcarbonyl (—CO—R) group,an alkoxycarbonyl (—CO—OR), an acyloxyalkyl-alkoxycarbonyl(—CO—O—R—O—CO—R) group where R is a monovalent or divalent group and asdefined above or a group having the formula —C(O)—O—CP1P2-haloalkyl,where P1 and P2 are the same or different and are H, lower alkyl, loweralkoxy, cyano, halo lower alkyl or aryl. Preferably the nitrogen atom isone of the nitrogen atoms of the amidino group of the compounds of theinvention. These prodrug compounds are prepared reacting the compoundsof the invention described above with an activated acyl compound to bonda nitrogen atom in the compound of the invention to the carbonyl of theactivated acyl compound. Suitable activated carbonyl compounds contain agood leaving group bonded to the carbonyl carbon and include acylhalides, acyl amines, acyl pyridinium salts, acyl alkoxides, inparticular acyl phenoxides such as p-nitrophenoxy acyl, dinitrophenoxyacyl, fluorophenoxy acyl, and difluorophenoxy acyl. The reactions aregenerally exothermic and are carried out in inert solvents at reducedtemperatures such as −78 to about 50C. The reactions are usually alsocarried out in the presence of an inorganic base such as potassiumcarbonate or sodium bicarbonate, or an organic base such as an amine,including pyridine, triethylamine, etc. One manner of preparing prodrugsis described in U.S. Ser. No. 08/843,369 filed Apr. 15, 1997(corresponding to PCT application WO9846576) the contents of which areincorporated herein by reference in their entirety.

Particularly preferred compounds of formula I and II are:

Synthesis

Compounds of the invention are prepared using standard organic synthetictechniques from commercially available starting materials such as thosedescribed in U.S. Pat. No. 6,469,047 which in its entirety isincorporated herein by reference. While various synthetic schemes can beemployed, the compounds of formula I in which L is —C(S)—O— may beprepared starting from a tyrosine ester according to the followingscheme

in which the starting tyrosine ester 1 is reacted with an acyl halide oracyl anhydride, e.g. acyl chloride 2, in THF with mild base, e.g. sodiumbicarbonate, to give intermediate 3. Intermediate 3 is then reacted withthiophosgene in methylenedichloride with diisopropylethylamine to givethiochloroformate 4 which is reacted with amine 5 in methylenedichlorideand diiodopropylethylamine to give thiocarbamate 6 ester. Conversion ofthe ester to a carboxylic acid is easily performed by saponificationwith an alkali-metal hydroxide such as lithium, sodium, or potassiumhydroxide. The starting tyrosine, acyl chloride intermediate 2 and amineintermediate 3 are either commercially available or are prepared fromstarting materials that commercially available employing establishedsynthetic techniques.

Numerous starting tyrosine derivatives are commercially available or canbe readily synthesized using standard chemical reactions. An example ofthe synthesis of a particular intermediate useful in preparing compoundsof the invention is:

In this scheme, R may be any suitable group which is non-reactive underthe reaction conditions. Examples of suitable R groups includesubstituted and unsubstituted alkyl, aryl, arylalkyl, etc. Additionalcompounds of the invention can then be prepared by acylating the phenylhydroxy group with an activated thiocarbonyl to form a thiocarbamate asdescribed herein.

Solid phase reaction chemistry provides a convenient method forsynthesizing the compounds of the invention. FMOC- or BOC-protectedamino acids and derivatives thereof are readily available and can beused as starting materials in the synthesis of the compounds of theinvention. The protected amino acid is initially attached to a syntheticresin having an available coupling group, such as an available hydroxy(e.g. benzyloxy resin beads). Coupling is achieved using known chemicalreactions, e.g. condensation reactions using for example DIPC or DMAP,to attach the amino acid to the solid support. Any known couplingreactions and resin surfaces may be used. The amino nitrogen is thendeprotected using, for example, a weak base such as piperidine or othersuitable base. The free amino group can then be reacted with anactivated ester such a HBTU or HOBT ester of a suitable group such as2-chlorobenzoic acid to form the desired N-substituted tyrosineintermediate.

Tyrosine intermediates are further reacted to form thiocarbamates, usingknown chemistry. For example, the hydroxy group can be reacted with athiocarbonyl synthon such as thiophosgene, followed by the desiredprimary or secondary amine, e.g. cyclic secondary amine, to formthiocarbamates as shown in the reaction scheme below.

In this scheme, a=DIPC cat./DMAP; b=20% piperidine/DMA or DMF; c=asubstituted benzoic acid/HBTU or other amide coupling agent/TEA or otherweak base; d=primary or secondary amine; e=TFA/triethylsilane, forexample.

Compounds of formula I may be synthesized manually via solid phasesynthesis on p-alkoxybenzyl alcohol resin (Advanced Chemtech, USA) asshown above. Commercially available FMOC protected tyrosine or othertyrosine analogs may be purchased from BACHEM Ca., Advanced ChemTechU.S.A., or Calbiochem Corp. (Ca.) or prepared from commerciallyavailable reagents. Typically 1 mmol of FMOC-tyrosine (or tyrosineanalog) is added to 1 g of p-alkoxybenzylalcohol resin in 50 mLdichloromethane. Diisopropylcarbodiimide (DIPC, 1 mmol) is addedfollowed by catalytic dimethylaminopyridine (DMAP, 0.1 mmol) and theresulting mixture is stirred under nitrogen at 20 C for 4 hours. Theresin is then washed with dichloromethane and dimethylacetamide (DMA)and the FMOC group is removed via mixing with 20% piperidine in DMA forfifteen minutes. The resin is then washed three times with DMA to removeexcess piperidine.

Ortho-chlorobenzoic acid (2 mmol) or other substituted benzoic acid ismixed with HBTU (2 mmol) or other suitable activating agent in 20 mL ofDMA and added to the previously washed resin. N-methylmorpholine ortriethylamine (4 mmol) is added and the mixture sparged with nitrogenfor 30 minutes. The resin is washed with dichloromethane and treatedwith 2 mmol of thiophosgene and 0.05 mmol DMAP in 20 mL of DMA for 1 h.Excess reagents are washed away and 2 mmol of morpholine or othersubstituted amine RaRb-NH in 20 mL dichloromethane is added. The mixtureis sparged overnight at room temperature and washed withdichloromethane.

Treatment with TFA containing 5% triethylsilane for 1 hour affords thecrude product. The crude material is extracted from the resin bystirring with 100 mL of 2:1H₂O/CH₃CN for 5 minutes followed byfiltration to remove the resin. The crude filtrate is lyophilized andpurified by preparative reverse phase C₁₈ HPLC (CH₃CN/H₂O gradient, 0.1%TFA) to afford purified material. Pure fractions are characterized byelectrospray ionization mass spectrometry (Sciex API100) and proton NMR,lyophilized to dryness and resuspended in DMSO at 10 mM just prior tobiological assay. Serial dilutions at 0.5 mM are titrated into an ELISAformat assay and the IC₅₀ for each compound may then be determined.

Alternatively, compounds of formula I can be synthesized in three stepsvia solution phase chemistry starting with commercially available(L)-tyrosine or tyrosine analogs having substituents at R². A generalsynthesis is depicted below. This type of synthesis is amenable to scaleup and for introducing ester prodrugs at R⁴.

Typically, 100 mmols of (L)-tyrosine or similar tyrosine analog isdissolved in 500 mL THF/H₂O (1:1) and 300 mmols of sodium bicarbonate isadded followed by 110 mmols (1.1 eq.) of a suitable benzoyl chloride oranhydride of general structure A-COCl. The solution is stirred at roomtemperature for 1 h. The mixture is concentrated via rotary evaporationand acidified to pH<3 with 1 N HCL. The acidified solution is extractedwith ethyl acetate and the organic layer is washed with satd. NaCl andevaporated to dryness. Crystallization of the crude material fromethylacetate/hexane affords pure compound as determined by analyticalHPLC.

If a suitable benzoyl chloride or anhydride is not available then thecorresponding substituted benzoic acid (100 mmols) is used incombination with HBTU or other amide coupling reagent. If this route isemployed, 100 mmols of (L)-tyrosine or similar tyrosine analog isdissolved in 250 mL of dimethylformamide. In a separate vessel, theappropriate benzoic acid (110 mmols) in DMF is mixed with 110 mmols ofHBTU or other amide coupling agent and 300 mmols of triethylamine orother weak base (NMM, DIPEA etc.). The mixture is allowed to stand for10 minutes and then added to the tyrosine in one portion. After stirringfor 1 hour at room temperature, the reaction mixture is concentratedunder high vacuum and resuspended in ethyl acetate. The suspension iswashed with 1 N HCL, water and satd. NaCl and evaporated to dryness.Crystallization affords pure compound.

A solution of thiophosgene (2 mmol) in 100 mL of dichloromethane iscooled to −78 deg under N₂. In a separate flask, N-substituted tyrosineintermediate (1 mmol) is dissolved in 20 mL of dichloromethane anddiisopropylethylamine (2 mmol) is added. The resulting mixture is addeddrop wise to the cooled thiophosgene via a syringe. The reaction isallowed to warm to room temp. and is stirred 2 h. The solvent is thenevaporated to dryness and the residue recrystallized from ethylacetate/hexaneto afford thiochloroformate.

The thiochloroformate intermediate (1 mmol) is dissolved indichloromethane (100 mL) and 2 mmol of morpholine or other organic amineis added. The reaction is stirred for 1 h at room temp. and evaporatedto dryness. The residue is dissolved in 1:1 acetonitrile/water and thethiocarbamate ester product purified by reverse phase HPLC(acetonitrile/water/0.1% trifluoroacetic acid).

The free alpha carboxylic acid may be converted to an ester or to anamide using reactions well known in the art. For example, a freecarboxyl group can be reacted with a suitable alcohol in the presence ofan acid to esterify the carboxyl group using well known reactions andreagents. Similarly, amides are formed by reacting the carboxylic acidwith an amine with removal of the water produced by the condensationusing known methods. A example of a reaction for esterification is shownbelow.

Compounds of formula I in which L is —C(O)—S— may be prepared by variousroutes using standard organic synthetic techniques from reagents thatare commercially available. In a particular scheme, such compounds aresynthesized in a similar manner to a carbamate starting with4-thiophenyl alanine amino acid analogs according to the followingscheme

in which the 5-thiophenyl alanine compound is reacted withR^(a)R^(b)N—C(O)X and excess base such as TEA, DIPEA, NMM, HCO₃— andOH—.Alpha-4 Inhibition

The compounds of the invention inhibit the binding of alpha-4 integrinsto their ligands, in particular alpha4beta1 and alpha4beta7 onlymphocytes, eosinophiles, basophiles and monocytes to a cell expressingVCAM-1 and/or MAdCAM on the cell surface. The inhibitory compounds ofthe invention are useful to prevent the interaction of an epithelialcell bearing VCAM-1 and/or MAdCAM on the cell surface with a leukocytecell bearing alpha4beta1 and/or alpha4beta7 on the surface by contactingthe epithelial cell or the leukocyte with an inhibitory amount of thecompound of the invention. The compounds are useful in assays todetermine the inhibitory effect of a compound which antagonizes thebinding of alpha4beta1 and/or alpha4beta7 integrin to VCAM-1 ligandand/or MAdCAM ligand. The inhibitory compound may be a small molecule, aprotein or peptide or an antibody. In an in vitro assay, the ligand orthe integrin may be directly or indirectly bound to a surface, such asmicrotiter plate, using known methods described for example in WO9820110, WO 9413312, WO 9624673, WO 9806248, WO 9936393, and WO 9910312.The other member of the binding pair, e.g. the integrin or the ligand,respectively, (or a cell expressing the same on its surface) is thenadded to the surface bound member and the inhibitory effect of a testmolecule is determined. The inhibitory activity of the compounds of theinvention can also be determined with this type of assay.

The binding of the integrins to their respective ligands is known to beinvolved in inflammatory conditions associated with leukocyteinfiltration of tissues lined with epithelial cells expressing VCAM-1 orMAdCAM. Such tissues include the gastrointestinal tract, skin, urinarytract, respiratory airways and joint synovial tissues. The compounds ofthe invention are useful in treating diseases in which such binding isimplicated as a cause of the disease or symptoms of the disease.Undesired disease symptoms may arise from cell adhesin and/or cellactivation which releases proinflammatory mediators, typically whenthere is an increase or upregulation in the expression of VCAM-1 and/orMAdCAM on the surface of endothelial cells. Various disease states whichcan be treated and for which the inflammatory symptoms can be reducedupon administration of the compounds of the invention include rheumatoidarthritis, asthma, psoriasis, multiple sclerosis, inflammatory boweldisease including ulcerative colitis, pouchitis and Crohn's disease,Celiac disease, nontropical Sprue, graft-versus-host disease,pancreatitis, insulin-dependent diabetes mellitus, mastitis,cholecystitis, pericholangitis, chronic sinusitis, chronic bronchitis,pneumonitis, collagen disease, eczema, and systemic lupus erythematosis.The compounds of the invention are useful in treating these diseases andconditions by inhibiting the integrin/ligand binding.

The compounds of the invention can be assayed for ability to block thealpha4beta7/MAdCAM-1 or alpha4beta1/VCAM-1 binding interaction byaddition of serial dilutions of the samples to plates with the receptorsas follows. 96-well plates are coated with mouse anti-human alpha-4(31470D, PharMingen, San Diego, Calif.). The plates are decanted andblocked with 0.5% BSA. After washing alpha₄beta₇ or alpha₄beta₁ isadded, followed by incubation for 2 h at room temperature. The platesare washed and samples of the small molecule antagonists are added tothe plates with MAdCAM-1-Ig-HRP or VCAM-1-Ig-HRP for 2 h at roomtemperature. After an additional wash, the bound MAdCAM-1-Ig-HRP orVCAM-1-Ig-HRP is detected by addition of tetramethylbenzidine (TMB,Kirkegaard & Perry, Gaithersberg, Md.), followed by detection of theabsorbance of the product.

Alternatively, the compounds can be assayed using any knownprotein-protein or cell-based assay method, such as those described, forexample, in WO 99/10312 (examples 179–180) and WO 99/36393 (RPMI-CS-1cell adhesion assay); Cardarelli et al., 1994, J. Biol. Chem.,269:18668–18673; and Viney et al, J. Immunol., 1996, 157: 2488–2497(cell adhesion assay).

For example, 96-well ELISA plates are coated overnight at 4° C. with 2μg/ml with anti-human CD49d, (31470D, PharMingen, San Diego, Calif.) inphosphate buffered saline. The plates are decanted and blocked withassay buffer (50 mM Tris-HCl, pH 7.5, 150 mM NaCl, 1 mM MnCl₂, 0.05%Tween-20 and 0.5% BSA) at room temperature for one hour, with gentleshaking. The plates are washed three times (in 50 mM Tris-HCl, pH 7.5,100 mM NaCl, 1 mM MnCl₂, 0.05% Tween-20) and 2 μg/ml of the desiredintegri (Genentech, Inc.) in assay buffer is added, followed byincubation at room temperature for two hours, with gentle shaking. Afterwashing three times, 50 μl of samples of the small molecule antagonists(serial dilutions from 10 mM stocks in 100% DMSO) are added to theplates with 50 μl of 1 μg/ml MAdCAM-1-Ig-HRP or VCAM-1-Ig-HRP(Genentech, Inc) in assay buffer. The plates are incubated two hours atroom temperature, with gentle shaking, followed by washing six times.The bound MAdCAM-1-Ig-HRP or VCAM-1-Ig-HRP is detected by addition ofthe peroxidase substrate, 3, 3′, 5, 5′, tetramethylbenzidine (TMB,Kirkegaard & Perry, Gaithersberg, Md.), for 10 minutes, followed byaddition of 1M phosphoric acid to stop the reaction. The absorbance ofthe solutions are read at 450 nm on a plate reader.

Suitable animal models exist for many diseases and conditions which canbe treated with the compounds of the invention. Additional confirmationof the efficacy of these compounds in specific diseases and at desireddoses can be assayed using these established models. For example, animalmodels of chronic inflammatory diseases such as asthma (Laberge, S. etal., Am. J. Respir. Crit. Care Med., 1995, 151:822–829.), rheumatoidarthritis (RA; Barbadillo, C. et al., Springer Semin. Immunopathol.,1995, 16:375–379), colitis (Viney et al, J. Immunol., 1996, 157:2488–2497) and inflammatory bowel diseases (IBD; Podalski, D. K., N.Eng. J. Med., 1991, 325:928–937; Powrie, F. et al., Ther. Immunol.,1995, 2:115–123) may be used to demonstrate the activity of thecompounds of the invention and to conduct dose and efficacy studies.

The invention also includes pharmaceutical compositions or medicamentscontaining the compounds of the invention and a therapeutically inertcarrier, diluent or excipient, as well as methods of using the compoundsof the invention to prepare such compositions and medicaments.Typically, the inhibitors used in the method of this invention areformulated by mixing at ambient temperature at the appropriate pH, andat the desired degree of purity, with physiologically acceptablecarriers, i.e., carriers that are non-toxic to recipients at the dosagesand concentrations employed into a galenical administration form. The pHof the formulation depends mainly on the particular use and theconcentration of compound, but preferably ranges anywhere from about 3to about 8. Formulation in an acetate buffer at pH 5 is a suitableembodiment.

The inhibitory compound for use herein is preferably sterile. Thecompound ordinarily will be stored as a solid composition, althoughlyophilized formulations or aqueous solutions are acceptable.

The composition of the invention will be formulated, dosed, andadministered in a fashion consistent with good medical practice. Factorsfor consideration in this context include the particular disorder beingtreated, the particular mammal being treated, the clinical condition ofthe individual patient, the cause of the disorder, the site of deliveryof the agent, the method of administration, the scheduling ofadministration, and other factors known to medical practitioners. The“effective amount” of the compound to be administered will be governedby such considerations, and is the minimum amount necessary to prevent,ameliorate, or treat the alpha-4 mediated disorder. Such amount ispreferably below the amount that is toxic to the host or renders thehost significantly more susceptible to severe infection.

As a general proposition, the initial pharmaceutically effective amountof the inhibitor administered parenterally per dose will be in the rangeof about 0.01–100 mg/kg, preferably about 0.1 to 20 mg/kg of patientbody weight per day, with the typical initial range of compound usedbeing 0.3 to 15 mg/kg/day. Oral unit dosage forms, such as tablets andcapsules, preferably contain from about 25 to about 1000 mg of thecompound of the invention.

The compound of the invention may be administered by any suitable means,including oral, topical, transdermal, parenteral, subcutaneous,intraperitoneal, intrapulmonary, and intranasal, and, if desired forlocal immunosuppressive treatment, intralesional administration.Parenteral infusions include intramuscular, intravenous, intraarterial,intraperitoneal, or subcutaneous administration.

An example of a suitable oral dosage form is a tablet containing 25 mg,50 mg, 100 mg, 250 mg, or 500 mg of the compound of the inventioncompounded with about 90–30 mg anhydrous lactose, about 540 mg sodiumcroscarmellose, about 5–30 mg polyvinylpyrrolidone (PVP) K30, and about1–10 mg magnesium stearate. The powdered ingredients are first mixedtogether and then mixed with a solution of the PVP. The resultingcomposition can be dried, granulated, mixed with the magnesium stearateand compressed to tablet form using conventional equipment. An aerosolformulation can be prepared by dissolving the compound, for example 5400mg, of the invention in a suitable buffer solution, e.g. a phosphatebuffer, adding a tonicifier, e.g. a salt such sodium chloride, ifdesired. The solution is typically filtered, e.g. using a 0.2 micronfilter, to remove impurities and contaminants.

EXAMPLES

The invention will be more fully understood by reference to thefollowing examples. They should not, however, be construed as limitingthe scope of the invention. All patent and literature citations areherein incorporated by reference in their entirety.

Example 1 Morpholino-Thiocarbamate Inhibitor

Synthesis of N-(2-chlorobenzoyl)-(L)-tyrosine ethyl ester (2)

Into a 2L round bottom flask was added 21 g (100 mmol) of (L)-tyrosineethyl ester (1) (Bachem) and 500 mL of tetrahydrofuran (THF). Themixture was magnetically stirred at room temperature until dissolved,and 500 mL of aqueous NaHCO₃ (0.5 M) was added. The mixture was cooledin an ice bath and 19.3 g (110 mmol) of 2-chlorobenzoyl chloride(Aldrich) was added slowly via a glass syringe. The ice bath was removedand the reaction was allowed to warm to room temperature with stirring.After 30 min., the reaction mixture was rotory evaporated under reducedpressure to remove the THF and the remaining aqueous bicarbonatesolution was extracted twice with 400 mL ethyl acetate. The combinedextracts were washed with 400 mL of satd. aqueous NaHCO₃, 400 mL of 0.1N HCl, 400 mL of water, and 400 mL of satd. aqueous NaCl. The remainingorganic layer was dried over Na₂SO₄ and evaporated to dryness to afford˜40 g of crude (2) as a pale yellow oil. The crude product (2) wasrecrystallized from ˜800 mL of boiling ethyl acetate/hexane (˜1:3) andcooled to 4° C. overnight. The resulting crystalline product wasfiltered and dried under a stream of nitrogen to afford 32 g (92%isolated yield) of purified (2). The N-(2-chlorobenzoyl)-(L)-tyrosineethyl ester product (2) was confirmed by NMR and electrospray massspectrometry (see attached) and judged >95% pure by HPLC and TLC(R_(f)=0.6, 1:1 ethyl acetate/hexane). The remaining 5% impurity(Rf=0.7) was identified as 2-chlorobenzoic acid by HPLC and TLC versusauthentic material.

Synthesis of Thiochloroformate Intermediate (3)

A solution of thiophosgene (2 mmol) in 100 mL of dichloromethane wascooled to −78 deg under N₂. In a separate flask,N-(2-chlorobenzoyl)-(L)-tyrosine ethyl ester (2) (1 mmol) was dissolvedin 20 mL of dichloromethane and diisopropylethylamine DIPEA (2 mmol) wasadded. The resulting mixture was added drop wise to the cooledthiophosgene via a syringe. The reaction was allowed to warm to roomtemp. and stirred 2 h. The solvent was evaporated to dryness and theresidue recrystallized from ethyl acetate/hexaneto affordthiochloroformate (3).

Synthesis of Thiocarbamate Ethyl Ester (4)

Thiochloroformate intermediate (3) (1 mmol) was dissolved indichloromethane (100 mL) and 2 mmol of morpholine or other organic aminewas added. The reaction was stirred for 1 h at room temp. and evaporatedto dryness. The residue was dissolved in 1:1 acetonitrile/water and thethiocarbamate ethyl ester product purified by reverse phase HPLC(acetonitrile/water/0.1% trifluoroacetic acid).

Synthesis of Morpholino-Thiocarbamate 5

Into a round bottom flask with a magnetic stirrer was added (0.65 mmol)of (4) (from above) and 30 mL THF. The mixture was stirred untildissolved and 20 mL of H₂O was added followed by 1 mL of aqueous 1M LiOH(1 mmol). The resulting suspension was stirred vigorously at roomtemperature for 2 h or until TLC (5% CH₃CO₂H in ethyl acetate) indicatedcomplete disappearance of starting material. The reaction mixture wasacidified to pH<3 via addition of 0.1 M aqueous HCl and concentrated viarotory evaporation to remove most of the THF. The resulting aqueoussuspension was extracted 2× with 40 mL of ethyl acetate. The combinedorganic extracts were dried over Na₂SO₄ and rotory evaporated to affordcrude (5). The crude product was recrystallized from ethylacetate/hexane (˜2:1) and cooled to 4 deg. overnight. The crystallineproduct was filtered and dried under a stream of nitrogen to affordmorpholino-thiocarbamate product (5) as a pale yellow solid.

In a similar manner compounds were produced in which morpholine wassubstituted with piperidine, piperazine and N-acetyl-piperazine.

Example 2 Pharmacokinetic Properties and Activity

Clearance and Half Life

Jugular Vein Cannulation—Animals are anesthetized via IP injection usingKetamine/Xylazine/saline solution (@ 0.25 mL/kg). Animals are weighedprior to dosing of anesthetic to determine proper dosage. Sterileinstruments and aseptic technique are used throughout surgery. Thisincludes wearing a mask, clean lab coat or scrubs and sterile gloves.The ventral and dorsal neck areas are shaved and prepped with betadineand alcohol. A small skin incision is made over the jugular vein. Usingblunt dissection techniques, free the intended vessel from surroundingtissue and thread two sutures under the vein. Tie the cranial suture,nick the vessel, insert the catheter, and use the distal suture tosecure the catheter. Dissect a subcutaneous passage between the catheterinsertion point and the intrascapular space; make a small exit hole atthe nape of the neck. Then, using hemastats, pull the cannula throughthe passage to the dorsal neck area. Confirm that the catheter is stillproperly placed, flush with appropriate heparin/saline solution, andknot the distal end of the cannula. Place a suture tie around the knot,coil the cannula under the skin and close the dorsal incision, leavingthe “tie” slightly exposed for ease of externalizing the catheter. Closethe ventral incision. The animal should be recovered on a circulatingheating blanket or equivalent and returned to its room when it's able toright itself.

Test Articles

Compounds are formulated with polyethylene glycol 400 (PEG) at 30% (IV)or 60% (PO).

Dose Administration

Intravenous (IV) dosing is accomplished with a bolus injection into alateral tail vein. Animals are restrained using a rat restrainer tominimize mis-dosings and to reduce animal stress. Individual doses arecalculated based on body weights taken the morning of the dose. Oral(PO) dosing is accomplished by oral gavage using a 3½ inch stainlesssteel animal feeding tube. Animals are restrained by grasping gentlywith our hands to reduce animal stress. Individual doses are calculatedbased on body weights taken the morning of the dose.

Blood Sample Collection

Blood (approximately 0.2 mL) is collected from an jugular cannula. Foroccasions when the jugular cannula fails, blood is removed from theremaining lateral tail vein. The whole blood was placed intoMicrotainer® tubes containing K₂EDTA anticoagulant. Samples are invertedseveral times to ensure proper mixing with anticoagulant and are storedon ice until centrifugation. Samples are centrifuged at 10,000×g for 5minutes and plasma is transferred to 1.5 mL microcentrifuge tube. Bloodsamples, for IV dose administrations are collected prior to the doseadministration (predose) and at 2, 5, 10, 20, 30, 45, 60, 120 minutes,4, 6, 8 and 10 hours postdose of the dose administration. For PO doseadministration, the blood collection time points are the same as IV doseadministration, except no blood sample is collected at 2 minutes.

All plasma samples are measured by LC/MS/MS. All pharmacokineticparameters, clearance (CL), half life (t1/2), area under curve (AUC) andmaximum conc (Cmax) are determined using WinNonin (version 3.2).

TABLE 1 compound Assay Result referencecarbamate

Cl (ml/min/kg) 50AUCmet/AUCpar 0.96T1/2 (min) 6 1

Cl (ml/min/kg) 23AUCmet/AUCpar 0.04T1/2 (min) 61

1. A method for treating an adverse inflammatory response mediated bythe binding of an alpha-4 integrin to an alpha-4 integrin ligandcomprising administering to a mammal in need thereof an effectiveinhibitory amount of the compound of formula I:

wherein q is 0 or 1; T is O, CHR⁶, NR⁶, S, SO, SO₂, —NR⁶C(O)—,—C(O)NR⁶—; R^(a) and R^(b) are each independently hydrogen, alkyl,alkoxy, a carbocycle, a heterocycle, optionally substituted withhalogen, hydroxy, amino, carboxyl, nitro, cyano, a carbocycle or aheterocycle; and one to three carbon atoms of said alkyl and alkoxygroups are optionally replaced with carbonyl, NR⁶, O, S, SO or SO₂; orR^(a) and R^(b) together with the nitrogen to which they are attachedmay form a heterocycle or heteroaryl group substituted with 0–4 R¹substituents; R^(c) is H, alkyl, optionally substituted with hydroxy,halogen, alkoxy, amino, a carbocycle or a heterocycle; and one to threecarbon atoms of said alkyl are optionally replaced with carbonyl, NR⁶,O, S, SO or SO₂; L is —C(S)—O— or —C(O)—S—; Y is CH₂ or absent when p is0; Z is H or lower alkyl, or when p is 1 then Z and Y together with theatoms from which they depend form a 5 member saturated or partiallyunsaturated 5 or 6 member heterocycle; R¹ in each instance isindependently selected from the group consisting of hydroxy, amino,amidine, guanidine, carboxyl, nitro, cyano, thiol, alkyl, alkoxy acarbocycle and a heterocycle wherein said alkyl and alkoxy groups areoptionally substituted with one or more hydroxyl, halogen, amino,amidine, guanidine, carboxyl, nitro, cyano, carbocycle or heterocycle;and one to three carbon atoms of said alkyl and alkoxy groups areoptionally replaced with carbonyl, NR⁶, O, S, SO or SO₂; and saidcarbocycle and heterocycle group is optionally substituted with one ormore hydroxyl, halogen, amino, amidine, guanidine carboxyl, nitro,cyano, alkyl, alkoxy or haloalkyl; or two R¹ substituents together withthe atoms from which they depend form a fused or bridged heterocycleoptionally substituted with one or more hydroxyl, halogen, amino,amidine, guanidine carboxyl, nitro, cyano, alky, alkoxy or haloalkyl; R²and R³ in each instance are independently selected from the groupconsisting of hydroxy, amino, amidine, guanidine, carboxyl, nitro,cyano, thiol, alkyl, alkoxy, a carbocycle and a heterocycle wherein saidalkyl and alkoxy groups are optionally substituted with one or morehydroxyl, halogen, amino, amidine, guanidine, carboxyl, nitro, cyano,alkoxy, carbocycle or heterocycle; and one to three carbon atoms of saidalkyl group is optionally replaced with carbonyl, NR⁶, O, S, SO or SO₂;and said carbocycle and heterocycle group is optionally substituted withone or more hydroxyl, halogen, amino, amidine, guanidine, carboxyl,nitro, cyano, alkyl, alkoxy or haloalkyl; R⁴ is H, alkyl, a carbocycleor heterocycle wherein said alkyl is optionally substituted with acarbocycle or heterocycle and said alkyl, carbocycle and heterocycle areoptionally substituted with lower alkyl, halogen, hydroxyl, alkoxy,haloalkyl or amino; R⁶ in each instance is independently H, alkyl or acarbocycle; m, n, and o are each independently 0–4; p is 0 or 1; andsalts and solvates thereof.
 2. A method for treating an adverseinflammatory response mediated by the binding of an alpha-4 integrin toan alpha-4 integrin ligand comprising administering to a mammal in needthereof an effective inhibitory amount of the compound of formula II:

wherein L is —C(S)—O— or —C(O)—S—; X is O, NR⁵, CR¹R⁶, S, SO or SO₂; Yis CH₂ or absent when p is 0; Z is H or lower alkyl, or when p is 1 thenZ and Y together with the atoms from which they depend form a 5 membersaturated or partially unsaturated 5 or 6 member heterocycle; R¹ in eachinstance is independently selected from the group consisting of hydroxy,amino, amidine, guanidine, carboxyl, nitro, cyano, thiol, alkyl, alkoxya carbocycle and a heterocycle wherein said alkyl and alkoxy groups areoptionally substituted with one or more hydroxyl, halogen, amino,amidine, guanidine, carboxyl, nitro, cyano, carbocycle or heterocycle;and one to three carbon atoms of said alkyl and alkoxy groups areoptionally replaced with carbonyl, NR⁶, O, S, SO or SO₂; and saidcarbocycle and heterocycle group is optionally substituted with one ormore hydroxyl, halogen, amino, amidine, guanidine carboxyl, nitro,cyano, alkyl, alkoxy or haloalkyl; or two R¹ substituents together withthe atoms from which they depend form a fused or bridged heterocycleoptionally substituted with one or more hydroxyl, halogen, amino,amidine, guanidine carboxyl, nitro, cyano, alky, alkoxy or haloalkyl; R²and R³ in each instance are independently selected from the groupconsisting of hydroxy, amino, amidine, guanidine, carboxyl, nitro,cyano, thiol, alkyl, alkoxy, a carbocycle and a heterocycle wherein saidalkyl and alkoxy groups are optionally substituted with one or morehydroxyl, halogen, amino, amidine, guanidine, carboxyl, nitro, cyano,alkoxy, carbocycle or heterocycle; and one to three carbon atoms of saidalkyl group is optionally replaced with carbonyl, NR⁶, O, S, SO or SO₂;and said carbocycle and heterocycle group is optionally substituted withone or more hydroxyl, halogen, amino, amidine, guanidine, carboxyl,nitro, cyano, alkyl, alkoxy or haloalkyl; R⁴ is H, alkyl, a carbocycleor heterocycle wherein said alkyl is optionally substituted with acarbocycle or heterocycle and said alkyl, carbocycle and heterocycle areoptionally substituted with lower alkyl, halogen, hydroxyl, alkoxy,haloalkyl or amino; R⁵ in each instance is independently selected fromthe group consisting of H, alkyl, a carbocycle and a heterocycle whereinsaid alkyl group is optionally substituted with one or more hydroxyl,halogen, amino, amidine, guanidine, carboxyl, nitro, cyano, alkoxycarbocycle or heterocycle; and one to three carbon atoms of said alkylgroup is optionally replaced with carbonyl, NR⁶, O, S, SO or SO₂; andsaid carbocycle and heterocycle group is optionally substituted with oneor more hydroxyl, halogen, amino, amidine, guanidine carboxyl, nitro,cyano, alkyl, alkoxy or haloalkyl; R⁶ in each instance is independentlyH, alkyl or a carbocycle; m, n, and o are each independently 0–4; p is 0or 1; and salts and solvates thereof.
 3. The method of claim 2, whereinX is O.
 4. The method of claim 2, wherein Y is absent and p is 0 and Zis H or alkyl.
 5. The method of claim 2, wherein R¹ in each instance isH; or alkyl, aryl, heteroaryl each optionally substituted with hydroxyl,halogen amino or cyano; or two R¹ substituents together with the atomsfrom which they depend form a fused aryl group or bridged heterocycleoptionally substituted with hydroxyl, halogen, amino, alky, alkoxy. 6.The method of claim 2, wherein X is O and R¹ is H, methyl or ethyl; ortwo R¹ substituents together with the atoms from which they depend forma 2-Oxa-5-azabicyclo[2.2.1]heptane bridged heterocycle.
 7. The method ofclaim 2, wherein X is NR⁵ wherein R⁵ is H, alkyl, alkanoyl, optionallysubstituted with hydroxyl, halogen, amino or aryl.
 8. The method ofclaim 7, wherein two R¹ substituents together with the atoms from whichthey depend form a 2,5-Diazabicyclo[2.2.1]heptane bridged heterocycle.9. The method of claim 2, wherein R² in each instance is independentlyhalogen, alkyl, alkoxy, aryl or aryloxy.
 10. The method of claim 9,wherein n is 1 and R² is adjacent to the thiocarbamate linkage and isselected from the group consisting of Cl, I, methyl, methoxy and phenyl.11. The method of claim 2, wherein R³ is halogen, nitro, carboxyl oralkylsulfonyl optionally substituted with halogen, hydroxyl or alkoxy.12. The method of claim 2, wherein R³ is Cl, nitro, carboxyl, —NHSO₂CF₃,—NHC(O)CF₃ or 4-methyl-phenyl.
 13. The method of claim 2, wherein R³ isCl in the ortho position and o is
 1. 14. The method of claim 2, whereinR⁴ is H, alkyl or aralkyl.
 15. The method of claim 2, wherein R⁴ is H.16. The method of claim 1, wherein R^(a) and R^(b) together with thenitrogen atom from which they depend form a group selected from thegroup consisting of


17. The method of claim 1, wherein R^(a) and R^(b) together with thenitrogen atom from which they depend form a group selected from thegroup consisting of


18. The method of claim 2, selected from the group consisting of:


19. The method of claim 1 wherein said adverse inflammatory response isrheumatoid arthritis, asthma, psoriasis, multiple sclerosis,inflammatory bowel disease, ulcerative colitis, pouchitis, Crohn'sdisease, Celiac disease, nontropical Sprue, graft-versus-host disease,pancreatitis, insulin-dependent diabetes mellitus, mastitis,cholecystitis, pericholangitis, chronic sinusitis, chronic bronchitis,pneumonitis, collagen disease, eczema or systemic lupus erythematosis.20. The method of claim 2 wherein said adverse inflammatory response isrheumatoid arthritis, asthma, psoriasis, multiple sclerosis,inflammatory bowel disease, ulcerative colitis, pouchitis, Crohn'sdisease, Celiac disease, nontropical Sprue, graft-versus-host disease,pancreatitis, insulin-dependent diabetes mellitus, mastitis,cholecystitis, pericholangitis, chronic sinusitis, chronic bronchitis,pneumonitis, collagen disease, eczema or systemic lupus erythematosis.